Protocol

Transcriptional Regulation

Volume 809 of the series Methods in Molecular Biology pp 321-333

Date:

Analysis of SUC2 Promoter Structure by Nucleosome Scanning

  • Jennifer ChangAffiliated withDepartment of Cell Biology, New York University School of Medicine
  • , Ales VancuraAffiliated withDepartment of Biological Sciences, St. John’s University Email author 

* Final gross prices may vary according to local VAT.

Get Access

Abstract

Chromatin remodeling is a key mode of transcriptional regulation, and studying the nucleosome positioning at promoters is an important means to understand how genes are regulated. Nucleosome scanning is a convenient method to study nucleosome positioning. Yeast cells are converted to spheroplasts and nuclei are isolated. The nuclei are then digested by micrococcal nuclease to yield mononucleosome-sized DNA. Using a set of overlapping primers that cover the entire promoter, quantitative real-time PCR is performed using the mononucleosome DNA as the template. The nucleosome enrichment for each primer is calculated to yield a map of nucleosome occupancy across the promoter.

Key words

Saccharomyces cerevisiae Nucleosome scanning Chromatin Promoter structure Transcription PLC1 Inositol polyphosphates